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Advanced method for cellular response analyses against drugs by simultaneous measurement of QCM and microscopy using the response model equations, and its applications

使用反应模型方程同时测量 QCM 和显微镜来进行药物细胞反应分析的先进方法及其应用

基本信息

批准号：
21K05118
负责人：
村松宏
金额：
$ 2.25万
依托单位：
Tokyo University of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2021
资助国家：
日本
起止时间：
2021-04-01 至 2023-03-31
项目状态：
已结题

项目摘要

今年度は、新たに５-フルオロウラシル（5-FU）、エトポシド（ETP）について、HepG2細胞で水晶振動子センサーでの共振周波数応答を調べた。5-FU, ETPのいずれについても、細胞への添加後に応答が現れるまでに、遅延時間があることがわかった。これは、5-FU, ETPともに作用が多段階で効果が生じることや、水への溶解度が比較的低く、設定できる濃度に制限があるためと推察した。遅延時間後の応答は、共に累積対数正規分布曲線でよく近似できることを確認した。水晶振動子センサーの共振周波数による細胞の形態変化の情報と細胞死の関係を調べるために、水晶振動子センサーによる測定と同時に、蛍光観察を行えるようにし、死細胞染色試薬のヨウ化プロピジウムを用いて、HepG2細胞にマイトマイシンC（MMC）を添加して測定を行った。細胞死が起きる条件のMMC濃度では、細胞の形態変化に対して死細胞の増加が遅れて生じることが示された。水晶振動子センサーと蛍光顕微鏡の同時測定によって、細胞の応答のより詳しい情報が得られることがわかった。補助事業期間全体として、水晶振動子センサーでの細胞の応答形式は、細胞接着課程では、一次遅れ応答、抗がん剤に対する応答では、低作用濃度で遅延時間があったのち累積対数分布曲線で近似でき、濃度の上昇に伴い、遅延時間が短縮・消失し、高作用濃度で、一次遅れ応答へと変化するモデル式で表されることが示された。この内容については、論文を投稿し、本報告書作成時点で修正版の審査中である。また、HepG2細胞に加えて、PC-3細胞、WI-38細胞、Hela細胞について、細胞接着応答を調べたところ、PC-3細胞は遊走性が高く、水晶振動子の共振周波数変化がほとんど得られない結果となった。WI-38細胞，Hela細胞も接着性が低く、HepG2細胞に比べて共振周波数の変化量が小さくなることがわかった。

This year, the new 5-FU, ETP and HepG2 cells were used to adjust the resonance frequency of crystal vibrators. 5-FU, ETP is not included in the list, the cell is added to the list, and the delay time is not included. 5-FU, ETP, etc., have a multi-stage effect, and the solubility of water is relatively low, and the concentration is limited. The normal distribution curve of the cumulative number of responses after delay time is approximately confirmed. The information of cell morphology and the relationship between cell death and resonance frequency of crystal oscillator were adjusted. The information of cell morphology and the relationship between cell death and resonance frequency of crystal oscillator were measured simultaneously. The information of cell morphology and the relationship between cell death and resonance frequency of crystal oscillator were measured simultaneously. The information of cell morphology and the relationship between cell death and resonance frequency of crystal oscillator were measured simultaneously. The information of cell morphology and HepG2 cell morphology and the relationship between cell death and resonance frequency of crystal oscillator were measured. The concentration of MMC in cell death and cell morphological changes are related to the increase of cell death and cell proliferation. The crystal oscillator is used for simultaneous measurement of the crystal structure and the cell structure. During the entire subsidy period, the response form of the cells in the crystal vibrator base is different from that of the cells, the response of the cells is different from that of the cells, the response of the cells is different from that of the anti-interference agents, the delay time at low effective concentrations is similar to that of the cumulative logarithmic distribution curve, the delay time is shortened and disappeared with the increase of concentration, and the response of the cells is different at high effective concentrations. The content of this report is in the process of being submitted, and the revised version is under review at the time of preparation of this report. HepG2 cells, PC-3 cells, WI-38 cells, Hela cells, cellular responses, PC-3 cells, high mobility, crystal vibrators resonance frequency changes. WI-38 cells, Hela cells and HepG2 cells have low adhesion and low resonance frequency.